ABSTRACT
The serodiagnosis of small ruminant lentivirus (SRLV) infections in goats and sheep is usually performed by the agar-gel immunodiffusion technique (AGID) and by ELISA. Therefore, the western blot (WB) is the choice for the definition of antigen recognition patterns during the disease progression, being potentially useful for the diagnosis itself. In the present study, the antigen used for WB was obtained by a novel simple purification procedure: the dialysis of the supernatant of infected cultured cells, followed by centrifugation in sucrose gradient. Proteins in the pellet were separated by SDS-PAGE at 10% density and transferred to nitrocellulose membranes in a semi-dry blotting device. After development, five viral proteins were recognized by the standard positive serum sample, with molecular weights of 14-16, 25, 40, 50 and 70 kDa respectively. All 8 AGID positive goat serum samples recognized at least one band in the immunoblot, with different intensities. The total number of bands recognized by each serum sample varied considerably. A positive reaction to gp40 was observed with 4 sera, although rather weak in 3 cases. Two samples were reactive to p16 and two others to gp50, although weakly. Curiously, no serum was reactive to the 70 kDa antigen, recognized by the standard positive serum sample. These results suggest that the WB can be effectively used for the routine serodiagnosis of viral caprine arthritis encephalitis infection (CAEV). It can also support further studies on antigen recognition patterns with the new antigen provided by the novel simplified purification system of viral components described here.
KEY WORDS
Cells; corneal; purification; SDS-PAGE; immunoblot
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